Principles of Fluorescence Spectroscopy

PRINCIPLES OF FLUORESCENCE SPECTROSCOPY 877
Figure I.4. Corrected and normalized fluorescence spectra for some
β-carboline derivatives: (1) harmine 0.1 N H 2 SO 4 ; (2) 2-methyl
harmine 0.1 N H 2 SO 4 ; (3) harmane 0.1 N H 2 SO 4 ; (4) nor-harmane 0.1
N H 2 SO 4 ; (5) harmaline 0.01 N H 2 SO 4 . Revised from [2]. Copyright
© 1992, with permission from Elsevier Science.
3. CORRECTED EMISSION SPECTRA OF
9,10-DIPHENYLANTHRACENE, QUININE,
AND FLUORESCEIN
Corrected spectra in quanta per wavelength interval I(λ)
were published for these three compounds 4 (Figure I.5 and
Table I.6). The emission spectrum for quinine was found to
be at somewhat shorter wavelengths than that published by
Melhuish. 5
4. LONG-WAVELENGTH STANDARDS
Corrected emission spectra in relative quanta per wavelength
interval were reported 4 for quinine sulfate (QS), 3-
Figure I.5. Corrected emission spectra in relative photons per wavelength
interval (I(λ)) for 9,10-diphenylanthracene, quinine sulfate,
and fluorescein. From [4].
Table I.5. Corrected Emission Intensities for β-Carboline
in 1.0 N H 2 SO 4 at 25°C a
Wave- Corrected Wave- Corrected
length (nm) intensity length (nm) intensity
380 0.001 510 0.417
390 0.010 520 0.327
400 0.068 530 0.255
410 0.243 540 0.193
420 0.509 550 0.143
430 0.795 560 0.107
440 0.971 570 0.082
450 1.000 580 0.059
460 0.977 590 0.044
470 0.912 600 0.034
480 0.810 610 0.025
490 0.687 620 0.019
500 0.540 630 0.011
a Excitation at 360 nm. From [3].
aminophthalimide (3-APT), and N,N-dimethylamino-mnitrobenzene
(N,N-DMAMB). Chemical structures are
shown in Figure I.6. These standards are useful because
they extend the wavelength range to 750 nm (Figure I.7).
Table I.6. Corrected Emission Spectra in Relative Quanta
per Wavelength Interval (from [4])
Quinine
sulfate a Fluorescein b DPA c
λ (nm) I(λ) λ (nm) I(λ) λ (nm) I (λ)
310 0 470 0 380 0
350 4 480 7 390 39
380 18 490 151 400 423
400 151 495 360 412 993
410 316 500 567 422 914
420 538 505 795 432 1000
430 735 510 950 440 882
440 888 512 1000 450 607
445 935 515 985 460 489
450 965 520 933 470 346
455 990 525 833 480 222
457.2 1000 530 733 490 150
400 998 540 533 500 103
465 979 550 417 550 4
470 951 560 333 600 0
475 916 570 233
480 871 580 167
490 733 600 83
500 616 620 42
520 408 640 17
550 171 650 8
600 19 670 0
650 3
700 0
a Quinine sulfate was in 1.0 N H2 SO 4 , excitation at 346.5.
b Fluorescein (Uranine) was in 0.1 N NaOH, excitation at 322 nm.
c 9,10-diphenylanthracene (DPA) was in benzene, excitation at 385 nm.